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MassChrom® Steroids in Serum/Plasma with Sample Clean Up Columns - LC-MS/MS

Order No.: 72072/C/RUO, for 96 tests
Parameters:
11-Deoxycorticosterone, 11-Deoxycortisol, 17α-Hydroxyprogesterone, 21-Deoxycortisol, Aldosterone, Androstenedione, Corticosterone, Cortisol, Cortisone, Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone sulfate (DHEA-S), Dihydrotestosterone (DHT), Estradiol, Progesterone, Testosterone

Optimised and extended method
15 Steroids – 2 panels – 1 analytical column
With Sample Clean Up Columns

Steroid Panel 1

Steroid Panel 2

Steroid Panel 1

Aldosterone

Corticosterone

Cortisol

11-Deoxycortisol

21-Deoxycortisol

Cortisone

Steroid Panel 2

11-Deoxycorticosterone

Androstenedione

Dehydroepiandrosterone (DHEA)

Dehydroepiandrosterone sulfate (DHEA-S)

Estradiol

Progesterone

17α-Hydroxyprogesterone

Testosterone

Dihydrotestosterone (DHT)

This MassChrom® Steroid kit allows the quantitative determination of 15 steroids in serum/plasma by LC-MS/MS. Sample preparation is the same for both steroid panels using an optimised and efficient solid phase extraction with individual SPE columns (Sample Clean Up Columns). The use of stable isotope-labelled internal standards for every single analyte ensures reproducible and reliable quantitation of the steroids. The method does not require a high-end LC-MS/MS system to be carried out.

Alternatively, there is also a method available using 96 SPE well plates (72072/96/RUO, 72072/480/RUO).

More Information
Method of Analysis LC-MS/MS
Number of Tests 96
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Specimen Serum/Plasma
Sample Preparation
  • Put Sample Clean Up Columns into tubes for sample preparation.
  • Pipette 0.8 ml Equilibration Reagent 1 into each Sample Clean Up Column, centrifuge and repeat with Equilibration Reagent 2.
  • Pipette 500 μl research sample/calibrator/MassCheck® control, 50 μl Internal Standard Mix and 450 μl Extraction Buffer into each Sample Clean Up Column, mix briefly (vortex) and centrifuge.
  • Add 2 x 0.7 ml Wash Buffer and centrifuge.
  • Put autosampler vials into tubes for sample preparation, then put Sample Clean Up Columns on top.
  • Add 1 ml Elution Buffer and centrifuge.
  • Evaporate eluates in the autosampler vials (nitrogen or compressed air) to dryness.
  • Add 100 μl Reconstitution Buffer.
  • Close autosampler vials, mix briefly (vortex) and inject 5–50 μl of the eluate.  
Run Time Steroid Panel 1: 10.0 min; Steroid Panel 2: 11.7 min
Injection Volume 5-50 µl
Column Temperature Steroid Panel 1: 32 °C, Steroid Panel 2: 27 °C
Gradient binary
Ionisation ESI negative and positive (ESI switch)
MS/MS Mode MRM
Parameters 11-Deoxycorticosterone, 11-Deoxycortisol, 17α-Hydroxyprogesterone, 21-Deoxycortisol, Aldosterone, Androstenedione, Corticosterone, Cortisol, Cortisone, Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone sulfate (DHEA-S), Dihydrotestosterone (DHT), Estradiol, Progesterone, Testosterone
The following components are included in the kit:
  1. Mobile Phase A
    Mobile Phase A
    Order no.: 72011/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  2. Mobile Phase B
    Mobile Phase B
    Order no.: 72002/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  3. 72033 Elution Buffer for Steroids in Serum Plasma
    Elution Buffer
    Order no.: 72033/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  4. 72044 internal standard mix steroids serum plasma
    Internal Standard Mix
    Order no.: 72044/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  5. Extraction Buffer
    Extraction Buffer
    Order no.: 72005/RUO

    MassChrom® Steroids in Serum/Plasma

    Details
  6. Reconstitution Buffer
    Reconstitution Buffer
    Order no.: 72006/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  7. Equilibration Buffer 1 for Steroids in Serum/Plasma
    Equilibration Reagent 1
    Order no.: 72077/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  8. Equilibration Reagent 2
    Equilibration Reagent 2
    Order no.: 72008/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  9. Rinsing Solution
    Rinsing Solution
    Order no.: 72009/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  10. Wash Buffer
    Wash Buffer
    Order no.: 72012/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  11. 72055 Sample Clean Up Columns
    Sample Clean Up Columns
    Order no.: 72055

    MassChrom® Steroids in Serum/Plasma (72072/C/RUO) - LC-MS/MS

    Details
  12. Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Order no.: J0602
    Details
  13. J0410 J0602
    PP Screw-on Caps, pierceable silicone/PTFE septa, 1.0 mm
    Order no.: J0410
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 72110 Analytical Column for Steroids in Serum/Plasma
    Analytical Column Steroids in Serum/Plasma - LC-MS/MS
    Order no.: 72110

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  2. 72038 multilevel calibrator steroids panel1 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 1
    Order no.: 72038
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  3. 72039 multilevel calibrator steroids panel2 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 2
    Order no.: 72039
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  4. 0341 0342 0343 MassCheck Steroids Serum Controls
    MassCheck® Steroid Panel 1 Serum Controls
    Order no.: 0341/0342/0343
    Details
  5. 0345 0346 0347 LCMS steroid panel2 controls serum plasma
    MassCheck® Steroid Panel 2 Serum Controls
    Order no.: 0345/0346/0347
    Details
  6. System Check Solution Steroid Panel 1
    System Check Solution Steroid Panel 1
    Order no.: 72088/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  7. System Check Solution Steroid Panel 2
    System Check Solution Steroid Panel 2
    Order no.: 72099/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  8. Tuning Mix Steroid Panel 1
    Tuning Mix Steroid Panel 1
    Order no.: 72019/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  9. Tuning Mix Steroid Panel 2
    Tuning Mix Steroid Panel 2
    Order no.: 72020/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  1. 72039 multilevel calibrator steroids panel2 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 2
    Order no.: 72039
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  2. 72038 multilevel calibrator steroids panel1 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 1
    Order no.: 72038
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  1. 0345 0346 0347 LCMS steroid panel2 controls serum plasma
    MassCheck® Steroid Panel 2 Serum Controls
    Order no.: 0345/0346/0347
    Details
  2. 0341 0342 0343 MassCheck Steroids Serum Controls
    MassCheck® Steroid Panel 1 Serum Controls
    Order no.: 0341/0342/0343
    Details
  1. 72110 Analytical Column for Steroids in Serum/Plasma
    Analytical Column Steroids in Serum/Plasma - LC-MS/MS
    Order no.: 72110

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  1. Tubes for Sample Preparation
    Tubes for Sample Preparation
    Order no.: 33007
    Details
  2. Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Order no.: J0602
    Details
  3. J0410 J0602
    PP Screw-on Caps, pierceable silicone/PTFE septa, 1.0 mm
    Order no.: J0410
    Details
  4. Sample Concentrator
    Sample Concentrator for Autosampler Vials
    Order no.: 42730

    Fast evaporation of solvents
    Light and compact unit for easy use in fume cupboards
    High-quality stainless-steel needles
    Accurate height control
    Chromsystems-adapted blocks available for specific assay requirements

    Details
  5. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  6. Stainless Steel Prefilter - 0.5 µm
    Stainless Steel Prefilter - 0.5 µm
    Order no.: 15071
    Details

Steroid Panel 1

Steroid Panel 2

Steroid Panel 1

Aldosterone

Corticosterone

Cortisol

11-Deoxycortisol

21-Deoxycortisol

Cortisone

Steroid Panel 2

11-Deoxycorticosterone

Androstenedione

Dehydroepiandrosterone (DHEA)

Dehydroepiandrosterone sulfate (DHEA-S)

Estradiol

Progesterone

17α-Hydroxyprogesterone

Testosterone

Dihydrotestosterone (DHT)

This MassChrom® Steroid kit allows the quantitative determination of 15 steroids in serum/plasma by LC-MS/MS. Sample preparation is the same for both steroid panels using an optimised and efficient solid phase extraction with individual SPE columns (Sample Clean Up Columns). The use of stable isotope-labelled internal standards for every single analyte ensures reproducible and reliable quantitation of the steroids. The method does not require a high-end LC-MS/MS system to be carried out.

Alternatively, there is also a method available using 96 SPE well plates (72072/96/RUO, 72072/480/RUO).

More Information
Method of Analysis LC-MS/MS
Number of Tests 96
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Specimen Serum/Plasma
Sample Preparation
  • Put Sample Clean Up Columns into tubes for sample preparation.
  • Pipette 0.8 ml Equilibration Reagent 1 into each Sample Clean Up Column, centrifuge and repeat with Equilibration Reagent 2.
  • Pipette 500 μl research sample/calibrator/MassCheck® control, 50 μl Internal Standard Mix and 450 μl Extraction Buffer into each Sample Clean Up Column, mix briefly (vortex) and centrifuge.
  • Add 2 x 0.7 ml Wash Buffer and centrifuge.
  • Put autosampler vials into tubes for sample preparation, then put Sample Clean Up Columns on top.
  • Add 1 ml Elution Buffer and centrifuge.
  • Evaporate eluates in the autosampler vials (nitrogen or compressed air) to dryness.
  • Add 100 μl Reconstitution Buffer.
  • Close autosampler vials, mix briefly (vortex) and inject 5–50 μl of the eluate.  
Run Time Steroid Panel 1: 10.0 min; Steroid Panel 2: 11.7 min
Injection Volume 5-50 µl
Column Temperature Steroid Panel 1: 32 °C, Steroid Panel 2: 27 °C
Gradient binary
Ionisation ESI negative and positive (ESI switch)
MS/MS Mode MRM
Parameters 11-Deoxycorticosterone, 11-Deoxycortisol, 17α-Hydroxyprogesterone, 21-Deoxycortisol, Aldosterone, Androstenedione, Corticosterone, Cortisol, Cortisone, Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone sulfate (DHEA-S), Dihydrotestosterone (DHT), Estradiol, Progesterone, Testosterone
The following components are included in the kit:
  1. Mobile Phase A
    Mobile Phase A
    Order no.: 72011/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  2. Mobile Phase B
    Mobile Phase B
    Order no.: 72002/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  3. 72033 Elution Buffer for Steroids in Serum Plasma
    Elution Buffer
    Order no.: 72033/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  4. 72044 internal standard mix steroids serum plasma
    Internal Standard Mix
    Order no.: 72044/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  5. Extraction Buffer
    Extraction Buffer
    Order no.: 72005/RUO

    MassChrom® Steroids in Serum/Plasma

    Details
  6. Reconstitution Buffer
    Reconstitution Buffer
    Order no.: 72006/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  7. Equilibration Buffer 1 for Steroids in Serum/Plasma
    Equilibration Reagent 1
    Order no.: 72077/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  8. Equilibration Reagent 2
    Equilibration Reagent 2
    Order no.: 72008/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  9. Rinsing Solution
    Rinsing Solution
    Order no.: 72009/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  10. Wash Buffer
    Wash Buffer
    Order no.: 72012/RUO

    MassChrom® Steroids in Serum/Plasma (72072/RUO) - LC-MS/MS

    Details
  11. 72055 Sample Clean Up Columns
    Sample Clean Up Columns
    Order no.: 72055

    MassChrom® Steroids in Serum/Plasma (72072/C/RUO) - LC-MS/MS

    Details
  12. Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Order no.: J0602
    Details
  13. J0410 J0602
    PP Screw-on Caps, pierceable silicone/PTFE septa, 1.0 mm
    Order no.: J0410
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 72110 Analytical Column for Steroids in Serum/Plasma
    Analytical Column Steroids in Serum/Plasma - LC-MS/MS
    Order no.: 72110

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  2. 72038 multilevel calibrator steroids panel1 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 1
    Order no.: 72038
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  3. 72039 multilevel calibrator steroids panel2 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 2
    Order no.: 72039
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  4. 0341 0342 0343 MassCheck Steroids Serum Controls
    MassCheck® Steroid Panel 1 Serum Controls
    Order no.: 0341/0342/0343
    Details
  5. 0345 0346 0347 LCMS steroid panel2 controls serum plasma
    MassCheck® Steroid Panel 2 Serum Controls
    Order no.: 0345/0346/0347
    Details
  6. System Check Solution Steroid Panel 1
    System Check Solution Steroid Panel 1
    Order no.: 72088/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  7. System Check Solution Steroid Panel 2
    System Check Solution Steroid Panel 2
    Order no.: 72099/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  8. Tuning Mix Steroid Panel 1
    Tuning Mix Steroid Panel 1
    Order no.: 72019/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  9. Tuning Mix Steroid Panel 2
    Tuning Mix Steroid Panel 2
    Order no.: 72020/RUO

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  1. 72039 multilevel calibrator steroids panel2 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 2
    Order no.: 72039
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  2. 72038 multilevel calibrator steroids panel1 serum plasma
    6PLUS1® Multilevel Serum Calibrator Set Steroid Panel 1
    Order no.: 72038
    MassChrom® Steroids in Serum/Plasma - LC-MS/MS
    Details
  1. 0345 0346 0347 LCMS steroid panel2 controls serum plasma
    MassCheck® Steroid Panel 2 Serum Controls
    Order no.: 0345/0346/0347
    Details
  2. 0341 0342 0343 MassCheck Steroids Serum Controls
    MassCheck® Steroid Panel 1 Serum Controls
    Order no.: 0341/0342/0343
    Details
  1. 72110 Analytical Column for Steroids in Serum/Plasma
    Analytical Column Steroids in Serum/Plasma - LC-MS/MS
    Order no.: 72110

    MassChrom® Steroids in Serum/Plasma - LC-MS/MS

    Details
  1. Tubes for Sample Preparation
    Tubes for Sample Preparation
    Order no.: 33007
    Details
  2. Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Autosampler Vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml
    Order no.: J0602
    Details
  3. J0410 J0602
    PP Screw-on Caps, pierceable silicone/PTFE septa, 1.0 mm
    Order no.: J0410
    Details
  4. Sample Concentrator
    Sample Concentrator for Autosampler Vials
    Order no.: 42730

    Fast evaporation of solvents
    Light and compact unit for easy use in fume cupboards
    High-quality stainless-steel needles
    Accurate height control
    Chromsystems-adapted blocks available for specific assay requirements

    Details
  5. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  6. Stainless Steel Prefilter - 0.5 µm
    Stainless Steel Prefilter - 0.5 µm
    Order no.: 15071
    Details